Obtaining an affinity support for phospholipase A2 purification

Authors

  • Frenkel Guisado-Bourzac Universidad de Oriente
  • Dolores Lázara Romero-Del-Sol Centro de Estudio de Proteínas (CEP), Facultad de Biología, Universidad de La Habana
  • José Manuel Guisán-Seijas Instituto de Catálisis y Petroleoquímica, CSIC, Madrid
  • Jorge González-Bacerio Centro de Estudio de Proteínas (CEP), Facultad de Biología, Universidad de La Habana
  • Joaquín Díaz Brito Centro de Estudio de Proteínas (CEP), Facultad de Biología, Universidad de La Habana
  • Alberto del Monte-Martínez Centro de Estudio de Proteínas (CEP), Facultad de Biología, Universidad de La Habana

Abstract

The synthesis of a glyoxyl-Sepharose support was achieved departing from Sepharose CL-4B firstly activated, oxidized and aminated. Amination process is done in order to obtain an amino gel that could join specific ligands for phospholipases A2 purification. The egg yolk phosphatidylcholine (ePC) was immobilized by covalent method and was controlled by phosphate determination in the departure material and in laundries. This support has the advantage of being able to be chemically modified achieving different affinity supports. The validity of the supports obtained was checked by the addition of chromatography fractions from the sea anemones Condylactis gigantea and Stichodactyla helianthus, and the known snake venom from Crotalus durisus terrificus with phospholipase A2 activity. The typical elution maximum corresponding was obtained. Phospholipase A2 activity was corroborated qualitatively by a TLC-based method after exposure to purified ePC and fluorogenic substrate 1-palmitoil–2-NBD-C12-PC.

Keywords: affinity chromatography, phospholipase A2, egg yolk phosphatidylcholine immobilization, sea anemone, amine gel. 

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Published

2016-05-26

How to Cite

Guisado-Bourzac, F., Romero-Del-Sol, D. L., Guisán-Seijas, J. M., González-Bacerio, J., Díaz Brito, J., & del Monte-Martínez, A. (2016). Obtaining an affinity support for phospholipase A2 purification. Revista Cubana De Química, 28(2), 595–609. Retrieved from https://cubanaquimica.uo.edu.cu/index.php/cq/article/view/1146

Issue

Vol. 28 No. 2 (2016)

Section

Artículos

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